ATCC medium: 1425 Sporomusa medium
NaCl....................................... 2.25 g
MgSO4 . 7H2O ............................... 0.5 g
NH4Cl ...................................... 0.5 g
K2HPO4 ................................... 348.0 mg
KH2PO4 ................................... 227.0 mg
CaCl2 . 2H2O .............................. 25.0 mg
FeSO4 . 7H2O ............................... 2.0 mg
NaHSeO3 ................................... 15.0 mcg
Vitamin Solution (see below).............. 10.0 ml
Trace Elements Solution SL-6 (see below)... 3.0 ml
Yeast extract.............................. 2.0 g
Casitone (BD 225930)....................... 2.0 g
Resazurin.................................. 1.0 mg
8% NaHCO3 (see below) ..................... 50.0 ml
10% Glycine Betaine (see below)........... 50.0 ml
3% L-Cysteine . HCl (see below)........... 10.0 ml
Distilled water.......................... 877.0 ml
Boil medium for 5 minutes and cool in ice to room temperature under a gas stream of 80% N2, 20% CO2. Add sodium bicarbonate solution and autoclave medium anaerobically at 121C for 15 minutes.
Add Glycine Betaine Solution to cooled sterile medium. Add Cysteine Solution at time of inoculation. Adjust final pH if necessary, to 7.0-7.2.
Vitamin Solution:
Biotin.......................2.0 mg
Folic acid...................2.0 mg
Pyridoxine hydrochloride....10.0 mg
Thiamine . HCl...............5.0 mg
Riboflavin...................5.0 mg
Nicotinic acid...............5.0 mg
Calcium D-(+)-pantothenate...5.0 mg
Vitamin B12..................0.1 mg
p-Aminobenzoic acid..........5.0 mg
Lipoic acid..................5.0 mg
Distilled water............100.0 ml
Trace Elements Solution SL-6:
ZnSO4 . 7H2O .................0.10 g
MnCl2 . 4H2O .................0.03 g
H3BO3 ........................0.3 g
CoCl2 . 6H2O .................0.2 g
CuCl2 . 2H2O .................0.01 g
NiCl2 . 6H2O .................0.02 g
Na2MoO4 . 2H2O................0.03 g
Distilled water..............1.0 L
Adjust final pH of Trace Elements Solution SL-6 to 3.4.
8% NaHCO3:
Freshly prepared, gassed under 80% N2, 20% CO2 for 20 minutes and added to partially reduced (boiled) basal medium to yield a final concentration of 0.4%.
10% Glycine Betaine:
Filter-sterilized, gassed under 80% N2, 20% CO2 and aseptically added to sterile, partially reduced medium to yield a final concentration of 0.5% substrate.
3% L-Cysteine .HCl:
Freshly prepared, filter-sterilized, gassed under N2 and added aseptically to sterile medium at time of inoculation to yield a final concentration of 0.03% reducing agent.